Abstract: Methacrylic acid and ethylene glycol dimethacrylate were respectively used as functional monomer and cross-linker to synthesize molecularly imprinted polymers (MIPs), for adsorption of glutathione (GSH), named GSH-MIPs. Evaluation of various polymers by static experiment indicated the optimum ratio of template to functional monomer was 1:4; the optimal concentration and pH of loading buffer were 5.00-6.50 mmol L-1 and 5.0, respectively. By Scatchard analysis, the GSH binding site was found in GSH-MIPs, with K d and Q max of 1.96 mmol L-1 and 47.19 μmol g-1 , respectively. The method validated the extraction of GSH from yeast cells samples with the enrichment rate of 76%Template and target information:glutathione, GSHAuthor keywords:Molecularly imprinted polymers, Glutathione, GSH-MIPs, Adsorption conditions, selectivity, Efficient extraction